Fig. 1: CHIKV nsP1 curvature sensing ability on nanostructures in live cells. | Nature Communications

Fig. 1: CHIKV nsP1 curvature sensing ability on nanostructures in live cells.

From: Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cells

Fig. 1

a Schematic of a spherule on the plasma membrane (PM). The neck displays saddle curvature: positive (+) in the x-z plane and negative (-) in the x-y plane. Two types of nanostructures were designed to recapitulate the 3D saddle curvature into 2D. b Illustration and SEM image of the nanostructures: both 600 nm in height and 2 μm in length. The nanobar is 250 nm wide (pink arrow indicates positive curvature), and the nanocross is 350 nm wide (blue arrow indicates negative curvature). c U2OS cells cultured on nanobars with CellMask-stained PM (magenta) and overexpressed eGFP-nsP1 (green). The cell boundary shown in brightfield. n = 3 repeats. d Average images and 3D surface plots of PM (n = 489 bars) and eGFP-nsP1 (n = 223 bars) distribution around nanobars. e The end-to-center ratio of PM (n = 995) and eGFP-nsP1 (n = 1005) on nanobars. Mean ± SD [standard deviation]. f Cells cultured on nanocrosses with CellMask-stained PM (magenta) and overexpressed eGFP-nsP1 (green). Yellow arrows indicate negatively curved areas. n = 3 repeats. g Average images of PM and eGFP-nsP1 distribution around nanocrosses (n = 33 crosses). h Kymograph plots of eGFP-nsP1 on two adjacent nanobars. i Time-lapse images of eGFP-nsP1 clusters on (yellow arrows) and off (pink arrows) nanobars over 270 s. j Mean square displacement (MSD) of eGFP-nsP1 clusters on (n = 14) and off (n = 12) the nanobars in 110 s. Mean ± SEM [standard error of the mean]. k The trajectory of nsP1 clusters on (n = 14) and off (n = 12) bars over 110 s. l Confocal images of fluorescence recovery after photobleaching (FRAP) test for PM (magenta) and eGFP-nsP1 (green) on nanobar over 120 s. Yellow dashed circles indicate the bleached area. n = 3 repeats. m Normalized fluorescence intensity plot of PM and eGFP-nsP1 within nanobar area from FRAP measurement (2 s interval). Mean ± SD. Statistical analysis: two-tailed unpaired t test. Scale bars: 2 μm. Source data are provided as a Source Data file.

Back to article page